June 4 - 6, 2012
Crowne Plaza Hotel
St. Louis, MO
Abstracts
Food Allergens, Session Chair: Dr. Eric Garber


  • ISABEL COMINO1, ANA REAL1, Mª DE LOURDES MORENO1, LAURA DE LORENZO2, HUGH CORNELL3, MIGUEL ÁNGEL LÓPEZ-CASADO4,
    FRANCISCO BARRO5, PEDRO LORITE6, Mª ISABEL TORRES6, CAROLINA SOUSA1 AND ÁNGEL CEBOLLA7
    1 Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Sevilla, Sevilla, Spain
    2 Department of Plant Molecular Genetics, National Center for Biotechnology (CNB-CSIC), Madrid, Spain
    3 School of Applied Sciences, RMIT University, Melbourne, Australia
    4 Hospital Virgen de las Nieves, Granada, Spain
    5 Institute for Sustainable Agriculture (IAS-CSIC), Alameda del Obispo, Córdoba, Spain
    6 Department of Experimental Biology, Campus Las Lagunillas, Jaén, Spain
    7 Biomedal S.L., Sevilla, Spain.

    Beyond analysis of gluten content in food: assessment of potential toxicity for celiac patients by novel immunological methods.

    Celiac disease is an autoimmune disorder defined as an alteration of the small intestine mucosa associated with a permanent intolerance to gluten from wheat, barley,
    rye and, maybe oat. Currently, the only effective treatment is to avoid ingestion of gluten immunotoxic peptides. The relative proportion of toxic gluten peptides in
    food may diverge depending on many situations (cereal varieties, hydrolysis, protein modifications, etc). The most reliable method to assess the safety of food and
    beverages for celiac patients should determine the potential net immunotoxicity. Monoclonal antibodies G12 and A1 against the most immunotoxic fragment, the
    gliadin 33-mer peptide, showed to react only with immunotoxic gluten. The reactivity of immunological T cells from celiac patients tightly correlated to the reactivity
    of anti-gliadin 33mer antibodies when the degree of gluten hydrolysis with detoxifying glutenases was monitored. Furthermore, the anti-gliadin-33mer immunoassays
    allowed to identify varieties of oats with no significant immunotoxicity.
    _______________________________________________________________________________________________________________________________
    ANGEL CEBOLLA, Scientific Director and CEO,
    Biomedal S.L. Avda. Americo Vespucio 5-4, 41092 Sevilla;
    Phone: (+34) 954081276,
    e-mail: acebolla@biomedal.com

  • RICHARD E. GOODMAN1, S.N. PRAMOD2, AFUA O. TETTEH1, LOTHAR VOGEL3, STEFAN VIETHS3.
    1 Food Allergy Research and Resource Program, Dept. of Food Science & Technology, University of Nebraska—Lincoln, 143 Food Industry Complex,        Lincoln,
    NE 68583-0955, USA
    2 Dept. of Biochemistry Sahyadri Science College, Kuvempu University, Shimoga, India
    3 Paul Ehrlich Institute, Langen, Germany

    Testing the biological relevance of human IgE binding to dietary antigens using a humanized rat basophil clone or IgE-stripped human peripheral blood
    basophils improves food safety evaluation.

    In vitro IgE binding assays are used to aid the diagnosis of allergies and in assessing potential risks of new dietary proteins introduced as novel food ingredients or
    through genetic engineering.  There are many reported examples of high-levels of antigen specific IgE binding to common dietary proteins with serum samples from
    subjects who do not experience food allergy to the food.  Cell based assays using rat basophil leukemia cells that were transfected with the gene for the FcεRI alpha-
    chain; or peripheral blood mononuclear cells from non-allergic donors that have been acid stripped of IgE can be sensitized with IgE from allergic subject’s serum and
    challenged with specific antigen to verify effective cross-linking and mediator release. Our assays demonstrated that sera with high IgE binding to common bean
    glycoproteins (phytohemagglutinin and alpha-amylase inhibitor) do not activate basophils and that these proteins are unlikely to elicit allergic reactions.
    _______________________________________________________________________________________________________________________________
    RICHARD E. GOODMAN, Research Professor,
    Food Allergy Research and Resource Program, Dept. of Food Science & Technology, University of Nebraska—Lincoln, 143 Food Industry Complex, Lincoln, NE
    68583-0955, USA;
    Phone: +1 (402) 472-0452,
    e-mail: rgoodman2@unl.edu

  • SIGRID HAAS-LAUTERBACH, MARKUS LACORN, ULRIKE IMMER
    R-Biopharm AG, An der neuen Bergstrasse 17, 64297 Darmstadt, Germany

    Measurement of protein hydrolysates by competitive  ELISAs: problems, solutions and limitations.

    After several years we now understand that allergens will never be measurable in a defined way like other analytes e.g. mycotoxins. By nature, allergens are nearly
    always a mixture of proteins with different mass fractions of each single protein. Sometimes these mixtures are used fractionated (whey proteins and caseins),
    changed during manufacturing (lactosylation of ß-lactoglobulin in skim milk powder) or denaturated by processing (beer, sourdough). This led us to the intuition that a
    standard reference material is not possible to obtain from a classical point of view as described in ISO standards. But, without such a material the starting point for
    every standardization effort is senseless, because results are not comparable. Until today, mainly all calibrators available for standardization of allergen test methods,
    are based on intact protein mixtures (e.g. NIST 8445, whole egg powder). For the detection of hydrolyzed or fragmented proteins, new calibrators closely related to
    the real conditions in processed food are necessary.
    _______________________________________________________________________________________________________________________________
    SIGRID HAAS-LAUTERBACH
    R-Biopharm AG, An der neuen Bergstrasse 17, 64297 Darmstadt, Germany
    Phone: +49 (0) 61 51 - 81 02 - 25
    e-mail: s.h.lauterbach@r-biopharm.de

  • JENNIFER A. VOYKSNER, ROBERT D. VOYKSNER
    LCMS Limited, 1502 W. NC 54, Suite 504, Durham, NC, USA, 27707.

    Discovery and Measurement of Physiologically Relevant Gluten Peptides from Wheat, Barley and Rye Cereal Grains using LC/MS.

    The Food Allergen Labeling and Consumer Protection Act of 2004 has identified eight foods as the most common food allergens. Wheat is one of these eight foods.
    Dietary gluten detection and quantification is important because some gluten proteins are implicated in a variety of immune diseases, food allergies and intolerances.

    New physiologically relevant gluten marker peptides have been identified in rye, barley and wheat. Our approach is designed for the discovery, identification and
    measurement of some of these peptides. Native cereal grains were proteolytically digested using conditions and enzymes that model the human digestion system and
    then treated with human tissue transglutaminase 2. A unique fluorescence tagging approach of the peptides, together with LC-MS/MS and LC-QTOF-MS, has enabled
    the discovery of more than 22 peptides that can serve as markers for the presence of wheat, rye and barley in foods and consumer products. Detection limits are in
    the low ppb range.
    _______________________________________________________________________________________________________________________________
    JENNIFER A. VOYKSNER
    LCMS Limited, 1502 W. NC 54, Suite 504, Durham, NC, USA, 27707
    Phone: 919-201-0051.
    e-mail: jennifer_voyksner@lcmslimited.com